血清对水牛卵母细胞体外成熟、体外受精及早期胚胎发育的影响

Effects of Serum on the Maturantion and Fertilization of Buffalo Oocytes as well as the Development of Early Buffalo Embryos in vitro

从屠宰场收集了本地水牛55头、黑白花奶牛22头的卵巢共获卵泡847枚,平均每个水牛卵巢回收3.67枚可用卵母细胞,约为黑白花奶牛(10.23枚/头)的1/3。试验分别采用添加与不添加血清的培养液体外成熟培养水牛卵母细胞,结果二者的成熟率无明显毒性差异(58.23%对56.67%),但体外受精后早期胚胎发育的8-细胞率有显著差异(35.4%对23.0%),表明体外成熟液中有无血清对水牛卵母细胞体外成熟率没有影响,但血清对卵母细胞的早期胚胎发育有重要影响。进一步比较成熟液中不添加血清但在受精液及胚胎液中添加血清和在各个阶段均有血清参与的早期胚胎发育率(8-细胞率),表明二者差异不显著(33.8%对35.4%)。经无血清成熟培养液培养的成熟卵母细胞可以经孤雌激活后得到早期胚胎(4细胞)。

847 oocytes from 55 local buffalo and 22 black-and-white milch cows were adopted in this experiment. The average quantity of usable oocytes from buffalo was much smaller than that from the cow （3.67 Vs 10.23）. It showed that the quality of buffalo ovary was poor. The addition of serum to culture medium for buffalo oocytes did not improved the maturation rate as compared to the medium without serum （58.23% Vs 56.67%）. However, serum did improve the 8-cell formation rate of early buffalo embryos as compared to the control without serum （35.4% Vs 23.0%）. The results indicated that serum has no effect on the in vitro maturation of buffalo oocytes while is advantageous to the development of early buffalo embryos in vitro. In order to confirm the role of serum, the oocytes cultured with or without serum both went through fertilization in medium with serum and then both of the early embryos were also cultured with serum, the results showed that the 8-cell formation rates of the two treatment were not different （35.4% Vs33.8%）. This experiment also demonstrated that the maturated oocytes in medium without serum could develop to 4-cell via parthenogenetic activation.